GSM7837631: CAL27_input; Homo sapiens; OTHER
ChIRP was carried out based on a standard ChIP assay using a SimpleChIP® Enzymatic Chromatin IP Kit (Cell Signaling Technology, USA). Briefly, cells were first transfected with biotinylated miRNA (RiboBio, China). Within 24 h, cells were prepared by crosslinking with formaldehyde. Chromatin was segmented using micrococcal nuclease, and cell lysis was carried out via sonication to break the nuclear membrane. Total DNA template (20 µL) was collected after centrifugation and sample dilution. The remaining segmented chromatin was then incubated with streptavidin magnetic beads (Merck, Germany) overnight at 4°C. The samples were collected with a magnetic stand and washed several times before elution. The eluted samples were reverse crosslinked and subjected to DNA purification. Finally, each sample DNA was eluted in a total volume of 30 µL twice and stored with templates at -20°C before sequencing. A total of 20 ng of DNA templates was used to build libraries for ChIRP-seq.